Sensitive determination of buspirone in serum by solid-phase extraction and two-dimensional high-performance liquid chromatography

A selective and sensitive determination of buspirone in serum by high-performance liquid chromatography is described. The procedure is based on separation on a C18 column. A solid-phase extraction procedure is used for sample clean-up. The retention on the first column is based on the hydrophobic interaction of buspirone with the stationary phase, and the retention on the second column is based on ionic interactions due to the presence of sodium lauryl sulphate in the mobile phase as well as hydrophobic interaction. This allows for good separation of buspirone from impurities and consequently allows lower detection limits than previously reported for liquid chromatographic methods. Detection by ultraviolet absorbance gives a detection limit of 0.2 ng/ml.     

On-line solid-phase extraction of piroxicam prior to its determination by high-performance liquid chromatography

A direct method for the determination of piroxicam in plasma is described. Plasma is directly injected onto the extraction column (10 mm x 2 mm I.D., packed with 40-microns Bond Elut C2) where proxicam is separated from the plasma concomitants using a solid-phase extraction procedure. Using a laboratory-made on-line column-switching system, the drug is quantitatively transferred and separated on the analytical column (15 cm x 4.6 mm I.D., Supelcosil LC18 DB, 5 microns) followed by determination using ultraviolet absorption at 331 nm. Validation of the method demonstrated a good recovery (100%), sensitivity (limit of determination 0.2 microgram/ml, based on a 20-microliters sample volume), accuracy and precision (better than 5%). The developed method has been adopted for studying the steady-state pharmacokinetics of the drug.     

An improved method for high-performance liquid chromatography of gossypol

Earlier liquid-chromatographic methods for the determination of gossypol, based on highly acidic methanolic solvents, provide broad tailing peaks. The use of acetonitrile with aqueous phosphoric acid/tri-n-butylammonium phosphate at pH 3.5 and a high-resolution radial-compression column gave greatly improved performance and excellent peak shape.     

An improved high-performance liquid chromatography method for the determination of homocysteine in human plasma

Summary Elevated plasma homocysteine is, a known risk factor in arteriosclerotic vascular disease. To measure homocysteine in a large number of samples, we have developed a rapid, simple, robust and inexpensive reversed-phase HPLC method for routine analysis. Mercaptopro-pionylglycine was used as the internal standard and an external calibration in plasma was performed. Improvement was achieved by the use of gradient elution (using a sodium acetate buffer and methanol) resulting in a higher number of samples analyzed per day. Plasma samples were reduced with tributylphosphine and the proteins were precipitated with perchloric acid before addition of internal standard. The analytes were derivatized by use of 7-fluorobenzofurazone-4-sulfonic acid ammonium salt. For calibration human plasma was spiked with nine different concentrations of homocysteine (range 2–50 μmol L⁻¹). The inter-assay precision of replicate (n=29) analysis of the concentration of homocysteine in a sample of pooled plasma was 3.0%. The limit of detection, defined as three times the signal-to-noise ratio, was 0.25 μmol L⁻¹. The linearity of the assay was confirmed for a plasma concentration range of 2–2000 μmol L⁻¹. The variation of duplicate analyses of 842 plasma samples was 2.6±1.7%.     

Simultaneous determination of five fat-soluble vitamins in feed by high-performance liquid chromatography following solid-phase extraction

A high-performance liquid chromatography method is developed for the simultaneous determination of menadione, retinyl acetate, cholecalciferol, alpha-tocopherol, and alpha-tocopherol acetate in feed. The present study uses an enzyme to destroy the coating film, ethanol to extract free vitamins, and Oasis HLB cartridges to purify. Vitamins are separated using an Atlantis dC18 column. The mobile phase is methanol-water (98:2 v/v). Detection is performed with a UV-vis detector at 230 and 265 nm. The linearity, accuracy, and repeatability of this method are all satisfactory. Application of the method is suitable for the determination of the fat-soluble vitamins in general feed.     

Study of solid-phase extraction for the determination of sequestering agents in river water by high-performance liquid chromatography

Anion-exchange solid-phase extraction accompanied with high-performance liquid chromatography has been developed for the determination of six kinds of aminopolycarboxylic acids (APCAs) in river water [N-(2-hydroxyethyl)ethylenediaminetriacetate (HEDTA), ethylenediaminetetraacetate (EDTA), 1,3-propanediaminetetraacetate (PDTA), diethylenetriaminepentaacetate (DTPA), 1,2-propanediaminetetraacetate (MeEDTA), and O,O′-bis(2-aminoethyl)ethyleneglycoltetraacetate (GEDTA)]. The enrichment of APCAs using an anion-exchange cartridge was successfully done by the removal of anions, which competed with APCAs in anion-exchange processes. Barium chloride solution was added to river water and the mixture was passed through On Guard II Ag and H cartridges and then a Bond Elut Jr.SAX cartridge to enrich APCAs. After elution, APCAs were analyzed on two reversed phase C30 columns connected in series and detected with ultraviolet detection. The enrichment using solid-phase extraction permitted the determination of APCAs in river water at concentrations as low as 1 nM. Good recoveries (83–111%) were obtained for each APCA by the standard addition method on three river water samples with high accuracy (RSD 1.8–9.5%). Applying this method, two kinds of APCAs, EDTA and DTPA, were determined in samples from the Oyabe and Senbo Rivers in Japan.     

On-line coupling of solid-phase extraction to high-performance liquid chromatography for determination of estrogens in environment

A method based on on-line solid-phase extraction (SPE) coupling to high-performance liquid chromatography (HPLC) for the determination of estrogens has been developed. This method can continuously perform extraction of estrone, estradiol, estriol and diethylstilbestrol from aqueous samples without any other pretreatment, which can then be analyzed by HPLC with a UV detector at 230 nm. A pre-concentration column was adapted with methanol/water for chromatographic separation and two kinds of sorbents were involved, which are octadecyl-bonded silica and cigarette filter. The condition of pH of samples, sample loading flow rate and desorption time were all optimized, and the performances of both two sorbents were satisfactory. The on-line SPE system requires very low maintenance and just involved a switching-valve-filter system and a flow-inject pump, and the operation of the whole SPE-HPLC instrumentation is quite simple. The detection limits for pre-concentrating 50 mL of standard solution using cigarette filter as sorbent ranged from 0.98 to 78.1 ng L⁻¹. The enhancement factors were in the range of 197-326. The recoveries of estrogens spiked in real water samples ranged from 85 to 112%. The precisions for nine replicate measurements of a standard mixture (5.0 μg L⁻¹) were in the range of 1.0-3.4%.     

Simultaneous determination of phenolic xenoestrogens by solid-phase extraction and high-performance liquid chromatography with fluorescence detection.

A highly sensitive and selective method for simultaneous determination of some hydroxyl group-containing endocrine disruptors, including bisphenol A (BPA), bisphenol B (BPB), bisphenol E (BPE), bisphenol F (BPF) and 4-nonylphenol (4-NP), was developed. The method consists of precolumn derivatization of the analytes, solid-phase extraction (SPE) and subsequent chromatographic analysis by high-performance liquid chromatography (HPLC) with fluorescence detection. 4,4'-Cyclohexylidenebisphenol (BPZ) was used as an internal standard. Derivatization was carried out using 4-(4,5-diphenyl-1H-imidazol-2-yl)benzoyl chloride (DIB-Cl) as a label. Parameters of the derivatization reaction (temperature, time, concentration of reagent, stability, etc.) and of the solid-phase extraction (recovery, solvent, etc.) were studied in detail. Detection limits of compounds studied in standard solutions ranged from 0.08-1.3 ppb (ng/ml). The proposed method was successfully applied to plastic samples; BPA was found in both polycarbonate and polyvinyl chloride plastics, while 4-NP was found in plastics made of polyvinyl chloride and another polymer.     

An improved method for cyanide determination in blood using solid-phase microextraction and gas chromatography/mass spectrometry

A new method is described for the qualitative and quantitative analysis of cyanide, a very short-acting and powerful toxic agent, in human whole blood. It involves the conversion of cyanide into hydrogen cyanide and its subsequent headspace solid-phase microextraction (HS-SPME) and detection by gas chromatography/mass spectrometry (GC/MS) in selected ion monitoring (SIM) mode. Optimizing the conditions for the GC/MS (type of column, injection conditions, temperature program) and SPME (choice of SPME fiber, effect of salts, adsorption and desorption times, adsorption temperature) led to the choice of a 75-microm carboxen/polydimethylsiloxane SPME fiber, with D3-acetonitrile as internal standard, and a capillary GC column with a polar stationary phase. Method validation was carried out in terms of linearity, precision and accuracy in both aqueous solutions and blood. The limit of detection (LOD) and limit of quantitation (LOQ) were determined only in aqueous solutions. The assay is linear over three orders of magnitude (water 0.01-10, blood 0.05-10 microg/mL); and the LOD and LOQ in water were 0.006 and 0.01 microg/mL, respectively. Good intra- and inter-assay precision was obtained, always <8%. The method is simple, fast and sensitive enough for the rapid diagnosis of cyanide intoxication in clinical and forensic toxicology.     

Determination of acetaldehyde in human blood by high-performance liquid chromatography using fluorometry

A method was developed for the measurement of acetaldehyde in human blood by high-performance liquid chromatography (HPLC). The method was based on a pre-column reaction; a fluorescent substance was formed by a coupling reaction between 2 mol of cyclohexan-1,3-dione and 1 mol of acetaldehyde with ammonium acetate. The coupling compound was analyzed by HPLC and the concentration of acetaldehyde was obtained from the calibration curve drawn from the results using standard solutions. The determination was sensitive and reproducible with a range of 0.2-10 microM and a precision (coefficient of variation) of 2.43%.     

Ion-pairing high-performance liquid chromatography determination of amitrole in apple after solid-phase extraction and precolumn derivatization

A novel method based on solid-phase extraction was studied for the extraction of amitrole (3-amino-1,2,4-triazole), and its residue determination in apples has been developed. The samples were derivatized with 4-chloro-3,5-dinitrobenzotrifluoride (CNBF). The derivatization conditions and the influence of elution composition on the separation were investigated. In pH 9.5 H₃BO₃–Na₂B₄O₇ media, the reaction of amitrole with CNBF was complete at 60°C after 30 min. The separation of derivatized amitrole was achieved at room temperature within 13 min by gradient elution mode with cetyltrimethylammonium bromide in mobile phase as ion-pair reagent. The method correlation coefficient was 0.9996, in concentrations ranging from 1.66 to 415 mg L⁻¹. The calculated recoveries of the proposed method were from 94.17% to 105.67%, and relative standard deviations were 1.57% to 6.44% in the application to the quantitative determination of amitrole in apples. The detection limit of amitrole was 0.10 mg L⁻¹ with a signal-to-noise ratio of 3. Figure Residue determination of amitrole in apple by ion-pairing high-performance liquid chromatography     

Determination of imazaquin residues in soil by solid-phase extraction and high-performance liquid chromatography

A method has been developed for the quantitation of imazaquin residues in soil. The herbicide was extracted from soil with methanol-water (2 + 1, v/v) and cleaned up by strong anion-exchange solid-phase extraction cartridges. Analysis was performed by using high-performance liquid chromatography with ultraviolet detection. Average recoveries through the method ranged from 90.7 to 100.6%, with relative standard deviation equal to or lower than 6.6%. The limit of detection was estimated to be 0.0015 mg/kg, and the minimum quantitation concentration of imazaquin in soil was 0.005 mg/kg. This method was successfully applied to evaluate imazaquin residue levels in soil and its dissipation rates in a soybean field in the Xisanqi District of Beijing, People's Republic of China. The dissipation study showed that the half life of imazaquin in soil was 10.37 +/- 0.0135 days at 3 different application rates.     

Urinary testosterone measurement by gas chromatography after solid-phase extraction and high-performance liquid chromatography.

A method was developed for the rapid determination of testosterone in urine. The procedure consists of solid-phase extraction (SPE) followed by high-performance liquid chromatographic (HPLC) clean-up before gas chromatographic determination. Recovery was evaluated by adding [3H]testosterone (10(4) cpm) to urine samples; the mean recovery of radioactivity after SPE and HPLC was 82%. Precision was estimated by repeated measurement of testosterone in four different urine samples; the coefficient of variation was 7.9% (95% confidence limits 6.1-11.4%). Accuracy was evaluated by standard addition and dilution assays; a linear relationship was found between the expected and observed values (r2 = 0.982). The method is rapid, effective and suitable for routine analysis.     

Optimization of high-performance liquid chromatography and solid-phase extraction for determination of organophosphorus pesticide residues in environmental samples

High-performance liquid chromatography with solid-phase extraction (HPLC-SPE) was optimized for the analysis of three organophosphorus pesticide residues in water, apples and vegetable samples. Octadecylsilica disks (47-mm diameter) were used for solid-phase extraction. The parameters that affect both separation and extraction of methyl parathion, parathion and phoxim, such as mobile-phase composition, ionic strength, temperature, pH, and breakthrough volume, were investigated. The application of optimized HPLC-SPE to environmental samples gave reproducible results with low detection limits of 5 µg L^?1 for methyl parathion and parathion and 2.5 µgL^?1. Precisions of less than 8, 9 and 12% were obtained for water, spinach and apple samples, respectively.     

分散固相萃取-高效液相色谱法测定水产品中7种麻醉剂

随着麻醉剂广泛用于渔业生产过程和水产品运输等领域,建立水产品中麻醉剂残留的检测方法具有重要意义.由于水产品基质复杂且麻醉剂残留水平低,因此需要合适的前处理方法以提高检测灵敏度.该研究基于分散固相萃取-高效液相色谱,建立了一种同时检测水产品中普鲁卡因、丁氧卡因、三卡因、丁香酚、甲基丁香酚、异丁香酚、甲基异丁香酚7种麻醉剂...     

Determination of forchlorfenuron residues in watermelon by solid-phase extraction and high-performance liquid chromatography

A method using solid-phase extraction for cleanup, followed by high-performance liquid chromatography with ultraviolet detection (HPLC/UV), was developed for the determination of forchlorfenuron residues in watermelon. The pesticide is extracted from the sample with acidic acetonitrile, and the extract is loaded onto a primary-secondary amine (PSA) column. The pesticide is eluted with acetone and determined by HPLC/UV. The PSA column was found to provide effective cleanup, removing the greatest number of sample matrix interferences. The acetonitrile extraction followed by the PSA cleanup provided recoveries of >95%, coefficients of variation (precision) of <10%, and sensitivity of 0.005 mg/kg, in agreement with the directives for method validation in residue analysis. The proposed method was successfully used to determine forchlorfenuron residue levels and dissipation rates in watermelon grown in an experimental greenhouse in Beijing, People's Republic of China.     

Sensitive method for the determination of the antiarrhythmic drug detajmium in serum by solid-phase extraction and high-performance liquid chromatography with fluorimetric detection.

The objective of this study was to develop a very sensitive and selective method for the determination of detajmium (4-3-diethylamino-2-hydroxypropyl-ajmaline), a sodium-channel-blocking drug with antiarrhythmic properties, in serum. A high-performance liquid chromatography (HPLC) method with solid-phase extraction and fluorimetric detection has been applied. Serum samples were diluted with phosphate buffer (pH 3.5) and the extraction of detajmium and ajmaline, which was used as an internal standard, was carried out with Oasis cartridges (Waters). The chromatographic separation was performed on a RP18 column. The limit of quantification for serum samples of detajmium was 1 ng/ml with good reproducibility (R.S.D. < 15%) and a linear response from 1 to 200 ng/ml. The described method is highly sensitive and specific for the determination of detajmium in serum of patients and volunteers.